Cytogenetic Findings in a Primary Adrenocortical Carcinoma
Janusz Limon, Paola Dal Cin, Surabhi Kakati, Robert P. Huben, Uma Rao, and Avery A. Sandberg
ABSTRACT Cytogenetic analysis of a primary adrenocortical carcinoma revealed clonal rearrange- ments of several autosomes and sex chromosomes. In all metaphases the following mark- er chromosomes were present: 4p+,t(3;12)(p14;p13),14q+, t(15;20)(p11;q11), t(5;18) (p13.3;p11.2), psu dic(18)t(18;3)(p11.39;p12), and psu dic(20)t(20;9)(q11.2;p11). The results are discussed in relation to the cytogenetic findings in other solid tumors, especially of the kidney.
INTRODUCTION
Epithelial tumors of the adrenal cortex occur at all ages [1]. Most of these tumors occur in childhood and are endocrinologically active and malignant [2]. Hormon- ally inactive primary carcinomas of the adrenal cortex usually can be found in the elderly, but are extremely rare [3]. Primary carcinomas of the adrenal cortex account for less than 0.2% of all cancer deaths [1]. Fifty percent of these tumors are non- functional and are found mainly in patients over the age of 40, without any signif- icant differences in the sexes [4]. Neoplasms of the adrenal cortex consist of aden- omas and carcinomas, and among the nonfunctional tumors. 75% are diagnosed as carcinomas [1, 4].
Metastases to the adrenal gland occur frequently, particularly from cancers of the lung, breast, stomach, and kidney. Occasionally a primary adrenocortical car- cinoma may be difficult to distinguish histologically from renal cell carcinoma. As far as we know, no chromosome analysis of adrenocortical carcinoma has been pub- lished to date. We present herein the results of such an analysis on a primary non- functional adrenocortical carcinoma.
CASE REPORT
A 66-year-old white male was admitted to our institution with the diagnosis of a left adrenal tumor, which was found during the course of a work-up for metastatic cancer of unknown origin. He was initially seen with a lytic lesion in the left shoul- der; subsequently, clinical investigations revealed the presence of the above tumor. The patient had an extensive metabolic work-up, which included cortisol and 17-
From Roswell Park Memorial Institute, Buffalo, NY.
Address requests for reprints to Dr. Avery A. Sandberg, Roswell Park Memorial Institute, 666 Elm Street, Buffalo, NY 14263. Received June 25, 1986; accepted August 25, 1986.
ketosteroid levels, as well as urine and plasma catecholamines. All results were within normal limits. His past medical history revealed two transurethral resections of the prostate, both for benign disease. In August 1985 the patient underwent a left thoracoabdominal radical adrenalectomy, and a specimen of this tumor was ana- lyzed cytogenetically. The final pathologic diagnosis was adrenocortical carcinoma (stage IV) of left adrenal gland with extension to periadrenal fat (Figs. 1 and 2). Several months later the tumor metastasized to the bones.
MATERIALS AND METHODS
An excised piece of nonnecrotic tumor was finely minced with scissors in a Petri dish, transferred into culture flasks containing growth medium supplemented with collagenase (200 U/ml) and incubated for 16 hours [5]. The culture medium used was McCoy’s 5A (modified) supplemented with 17% fetal calf serum, L-glutamine, and antibiotics. Primary cultures were harvested after 48 and 72 hours and col- cemid was added in a final concentration of 0.01 µg/ml for 2-4 hours before har- vest. The cells were exposed to hypotonic shock according to Gibas et al. [6] and fixed with a mixture of methanol/glacial acetic acid (3:1) and dropped on cold, wet slides. GTW-banding was performed according to the method of Yunis [7] and CBG- banding according to that of Sumner [8].
Figure 3 GTW-banded karyotype from the adrenocortical carcinoma showing both numeri- cal and structural abnormalities: 54,XY, +X, +Y,-4,-4, - 10, -14, - 14, - 18, - 18, - 20, -20,- 22, +3, +5, +8, +19, +4p +, +4p +, + der(12)t(3;12)(p14;p13) +der(12)t(3;12)(p14;p13), +14q+, +14q+, + der(15)t(15;20)(p11;q11), + der(15)t(15;20)(p11;q11), + der(18)t(5;18) (p13.3;p11.2), +psu dic t(18;3)(p11.32;p12), +psu dic t(20;9)(q11.2;p11), +psu dic t(20;9)(q11.2;p11).
1
2
3
M1 MI
5
6
7
8
9
10
11
12
M2 M2
13 M3M3 15 M4M
16
17
MS M6
-
-
19
M7 M7
21
22
XX YY
RESULTS
Thirty two metaphases were analyzed; the modal chromosome number was found to be 54 (70% of metaphases). A duplication of the sex chromosomes was consis- tently present. Chromosomes #3, #5, #8, and #19 were trisomic in all cells and chromosome #16 in seven cells. Monosomy 10 and 22 was observed in all meta- phases (Fig. 2). Seven marker chromosomes were consistently present in all cells. The origin of most of these markers could be established as:
M1:4p+ or der(4)t(4;22)(p16;q11)
M2:der(12)t(3;12)(p14;p13)
M3:14q +
M4:der(15)t(15;20)(p11;q11)
M5:der(18)t(5;18)(p13.3;p11.2)
M6:psu dic(18)t(18;3)(p11.32;p12)
M7:psu dic(20)t(20;9)(q11.2;p11)
Structure of M6 and M7 was further supported by C-banding analysis. Both these chromosomes had two C bands (Fig. 3). Markers M5 and M6 existed in single copy; all other markers were present in two copies. In some metaphases additional mark- ers were found: t(7;7)(q36;q36) in two metaphases, t(15q15q) in two metaphases, and del(16)(q22) in three metaphases. Three metaphases with very high chromo- some numbers (98-108 chromosomes) were seen.
A
C
B
D
M6
M7
DISCUSSION
The cytogenetic results in an adrenocortical carcinoma obtained by us included stable clonal abnormalities involving chromosomes #3, #4, #5, #12, #14, #15, #18, and #20. A comparison of rearrangements of these chromosomes with those in urinary tract cancers revealed that some breakpoints involved in translocations were of interest, e.g., chromosome region 3p14 involved in the t(3;12). The same breakpoint, but in a t[3;11) in tumor cells, was found in a case from a family with hereditary renal adenocarcinoma [9]. Other investigators have described an associ- ation between constitutional chromosomal rearrangements t(3;8)(p14.2;q24.13) with hereditary renal carcinoma [10-12]. Furthermore, abnormalities of the 3p re- gion have been described in nonfamilial renal cell carcinoma [13, 14]. Thus, alter- ations in 3p may be possibly associated with the development of renal cell carci- noma [9, 12-14]. On the other hand, chromosome #3 is frequently affected in both hematologic malignancies and in some solid tumors (e.g., small cell lung carci- noma) [15]. Therefore, some investigators have suggested a more general role for this chromosome alteration in malignancy [16]. Recently, two oncogenes have been localized to the involved part of chromosome #3: c-raf-1 and tentatively c-H-ras [17, 18]. Thus, involvement of this chromosome region appears to indicate a rela- tionship between a specific chromosome alteration in cancer cells, location of cer- tain oncogenes, and existence of fragile sites. Among the structural rearrangements in our case, chromosome #5 was involved in a complex rearrangement forming marker M5 [i.e., t(5;18)(p13.3;p11.2)]. The same chromosome was found to be in- volved in two kidney tumors [14, 19], but the break occurred at band q13 and the distal part was translocated to chromosome #14.
Our case of adrenocortical carcinoma presented some unique karyotypic fea- tures. Although chromosome #3 is involved in rearrangements in renal tumors, other chromosome aberrations found in our case have not been described in renal or bladder tumors. For example, psu dic t(20;9), M7, was present in the tumor cells of only one of 44 cases of ovarian cancer [20]. The finding of recurring aberrations in adrenal tumors may allow us to differentiate them from others, especially clear cell carcinoma, help in the diagnosis of these tumors, and point to the origin of metastatic lesions whose primary origin cannot be readily ascertained.
The authors express their thanks to Eugenia Pietrzak for skillful technical assistance and to Anne Marie Dulinawka and Sharon Zolnowski for clerical assistance.
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